阿嗪米特原料药中潜在基因毒性杂质氯乙酰氯和氯乙酸的含量测定
Determination of potential genotoxic impurities chloroacetyl chloride and chloroacetic acid in azintamide raw material
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摘要: 采用化学衍生化-高效液相色谱法分别测定阿嗪米特原料药中潜在基因毒性杂质氯乙酰氯和氯乙酸。以2-硝基苯肼为衍生化试剂，经衍生化后以Thermo syncronis C18（250 mm × 4.6 mm，5 μm）为色谱柱，以0.1%磷酸水-乙腈为流动相进行梯度洗脱；柱温40 ℃，检测波长226 nm；流速1 mL/min。空白溶剂、衍生化试剂及阿嗪米特均不干扰待测物质出峰；氯乙酰氯和氯乙酸的检测限分别为7.5 ng/mL和15 ng/mL，均在30 ~ 300 ng/mL的范围内与色谱峰面积呈现良好的线性关系；加样回收率在87.37% ~ 109.75%范围内。本研究所建立的两种方法专属性好、精密度好、灵敏度高、操作简便，可用于阿嗪米特原料药中潜在基因毒性杂质氯乙酰氯及氯乙酸的痕量测定。Abstract: In this paper, chemical derivatization-high performance liquid chromatography was used to determine the potential genotoxic impurities chloroacetyl chloride and chloroacetic acid, respectively, in the raw material of azintamide.Derivatization was carried out using 2-nitrophenylhydrazine followed by the determination.Separation was performed on a Thermo Syncronis C18 column (250 mm × 4.6 mm, 5 μm), with mobile phase consisting of 0.1% phosphoric acid in water (A) and acetonitrile(B) by gradient elution, at a flow rate of 1 mL/min.The column temperature was 40 °C and the detection wavelength was 226 nm.The blank solvent, derivatization reagent, and azintamide did not interfere with the peak of the test substance, and the target component was well separated from the others.For impurities chloroacetyl chloride and chloroacetic acid, the limits of detection (LOD) were 7.5 ng/mL and 15 ng/mL respectively. There was a good linear relationship between the integral area and the concentration in the range of 30-300 ng/mL.The sample recovery rate was in the range of 87.37% ~ 109.75%.The two methods established in this study have good specificity, good precision, high sensitivity and simple operation, which can be used for the trace determination of potential genotoxic impurities chloroacetyl chloride and chloroacetic acid in the raw material of azintamide.
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Keywords:
- azintamide /
- genotoxic impurity /
- HPLC /
- pre-column derivatization /
- chloroacetyl chloride /
- chloroacetic acid
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CH M7（R1）：Assessment and control of DNA reactive（mutagenic）impurites in pharmaceuticals to limit potential carcinogenic risk
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